Inhibitors of matrix metallaproteinases

ABSTRACT

The invention provides compounds that inhibit MMPs; methods for treating or preventing cancer, angiogenesis, arthritis, connective tissue disease, cardiovascular disease, inflammation or autoimmune disease in a mammal; a method for inhibiting a matrix metalloproteinase in vivo or in vitro; and a method for imaging a tumor in vivo or in vitro.

RELATED APPLICATION

The present application claims priority under 35 U.S.C. 119 to U.S.Provisional Application Ser. No. 60/207,874; filed May 30, 2000 and U.S.Provisional Application Ser. No. 60/226,858; filed August 22, 2000;which applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

Specific interactions of cells within the extracellular matrix arecritical for the normal function of organisms. Alterations of theextracellular matrix are carried out by a family of zinc-dependentendopeptidases called matrix metalloproteinases (MMPs). The alterationsare carried out in various cellular processes such as organ development,ovulation, fetus implantation in the uterus, embyiogenesis, woundhealing, and angiogenesis. Massova, I.; Kotra, L. P.; Fridman, R.;Mobashery, S. FASEB J 1998, 12, 1075; Forget, M. -A.; Desrosier, R. R.;Béliveau, R. Can. J Physiol. Pharmacol. 1999, 77, 465-480.

MMPs consist of five major groups of enzymes: gelatinases, collagenases,stromelysins, membrane-type MMPs, and matrilysins. The activities ofMMPs in normal tissue functions is strictly regulated by a series ofcomplicated zymogen activation processes and inhibition by proteintissue inhibitors for matrix metalloproteinases (“TIMPs”). Forget, M.-A.; Desrosier, R. R.; Béliveau, R. Can. J. Physiol. Pharmacol. 1999,77, 465-480; Brew, K.; Dinakarpandian, D.; Nagase, H. Biochim. Biophys.Acta 2000, 1477, 267-283. Westermarck, J.; Kahari, V. M. FASEB J. 1999,13, 781-792. Excessive MMP activity, when the regulation process fails,has been implicated in cancer growth, tumor metastasis, angiogenesis intumors, arthritis and connective tissue diseases, cardiovasculardisease, inflammation and autoimmune diseases. Massova, I.; Kotra, L.P.; Fridman, R.; Mobashery, S. FASEB J. 1998, 12, 1075; Forget, M. -A.;Desrosier, R. R.; Béliveau, R. Can. J Physiol. Pharmocol. 1999, 77,465-480; Nelson, A. R.; Fingleton, B.; Rothenberg, M. L.; Matrisian, L.M. J. Clin. Oncol. 2000, 18, 1135.

Increased levels of activity for the human gelatinases MMP-2 and MMP-9have been implicated in the process of tumor metastasis. Dalberg, K.;Eriksson, E.; Enberg, U.; Kjellman, M.; Backdahl, M. World J. Surg.2000, 24, 334-340. Salo, T.; Liotta, L. A.; Tryggvason, K. J BioL Chem.1983, 258, 3058-3063. Pyke, C.; Ralfkiaer, E.; Huhtala, P.; Hurskainen,T.; Dano, K.; Tryggvason, K. Cancer Res. 1992, 52, 1336-1341. Dumas, V.;Kanitakis, J.; Charvat, S.; Euvrard, S.; Faure, M.; Claudy, A.Anticancer Res. 1999, 19, 2929-2938. As a result, select inhibitors ofMMPs (e.g., MMP-2 and MMP-9) are highly sought.

Several competitive inhibitors of MMPs are currently known. Theseinhibitors of MMPs take advantage of chelation to the active site zincfor inhibition of activity. Because of this general property, thesecompetitive inhibitors for MMPs are often toxic to the host, which hasbeen a major impediment in their clinical use. Greenwald, R. A. Ann. N.Y Acad. Sci. 1999, 878, 413-419; (a) Michaelides, M. R.; Curtin, M. L.Curr. Pharm. Des. 1999, 5, 787-819. (b) Beckett, R. P.; Davidson, A. H.;Drummond, A. H.; Huxley, P.; Whittaker, M. Drug Disc. Today 1996, 1,16-26.

Gelatinases have been shown to function in both female ovulation andinplantation of zygotes in the womb. The female contains a pair ofgonads, a system of ducts and chambers to conduct the gametes as well asto house the embryo and fetus, and external genitalia that facilitatereproductive function. The female gonads, the ovaries, lie in theabdominal cavity below most of the digestive system. Each ovary isenclosed in a tough protective capsule and contains many follicles. Afollicle consists of one egg cell surrounded by one or more layers offollicle cells, which nourish and protect the developing egg cell. Allof the 400,000 follicles a woman will ever have are formed at birth. Ofthese, only several hundred will be released during the woman'sreproductive years. After puberty, one (or rarely two or more) folliclematures and releases its egg during each menstrual cycle. The cells ofthe follicle also produce the primary female sex hormones, the estrogen.When ovulation occurs, the egg is expelled from the follicle (much likea small volcano), and the remaining follicular tissue grows within theovary to form a solid mass called the corpus luteum. The corpus luteumsecretes progesterone, the hormone of pregnancy, and additionalestrogen. If the egg is not fertilized, the corpus luteum degeneratesand a new follicle matures during the next cycle.

The female reproductive system is not completely closed, and the eggcell is expelled into the abdominal cavity near the opening of theoviduct, or fallopian tube. The oviduct has a funnellike opening, andcilia on the inner epithelium lining the duct help collect the egg cellby drawing fluid from the body cavity into the duct. The cilia alsoconvey the egg cell down the duct to the uterus, commonly called thewomb. The uterus is a thick, muscular organ shaped much like anupside-down pear. It is remarkably small; the uterus of a woman who hasnever been pregnant is about 7 cm long and 4-5 cm wide at its widestpoint. The unique arrangement of muscles that make up the bulk of theuterine wall allow it to expand to accommodate a 4-kg fetus. The innerlining of the uterus, the endometrium, is richly supplied with bloodvessels.

The pattern of hormone secretion controlling female reproduction differsstrikingly from the male pattern, reflecting a cyclic nature of femalereproduction.

Two different types of cycles occur in female mammals. Humans and manyother primates have menstrual cycles, whereas other mammals have estrouscycles. In both cases, ovulation occurs at a time in the cycle after theendometrium has started to thicken and become more extensivelyvascularized, which prepares the uterus for the possible implantation ofan embryo.

The menstrual cycle averages 28 days, but only about 30% of women havecycle lengths within a day or two of the statistical 28 days. Cyclesvary from one woman to another, ranging from about 20 to 40 days. Insome women the cycles are usually very regular, but in other individualsthe timing varies from cycle to cycle.

Paralleling the menstrual cycle is an ovarian cycle. It begins with thefollicular phase, during which several follicles in the ovary begin togrow. The egg cell enlarges and the coat of follicle cells becomesmulti-layered. Of the several follicles that start to grow, only oneusually continues to enlarge and mature, while the others degenerate.The maturing follicle develops an internal fluid-filled cavity and growsvery large, forming a bulge near the surface of the ovary. Thefollicular phase ends with ovulation when the follicle and adjacent wallof the ovary rupture, releasing the egg cell. The follicular tissue thatremains in the ovary after ovulation is transformed into the corpusluteum, an endocrine tissue that secretes female hormones during what iscalled the luteal phase of the ovarian cycle. The next cycle begins witha new growth of follicles.

Contraception literally means “against taking,” in this case, the takingin of a child. The term has come to mean preventing a pregnancy throughone of several methods. These methods fall into three main categories:(1) preventing the egg and sperm from meeting in the female reproductivetract, (2) preventing implantation of a zygote, and (3) preventing therelease of mature eggs and sperm from the gonads.

Besides complete abstinence, the methods that prevent release of gametesare the most effective means of birth control. Chemical contraception(birth control pills) have failure rates of less than 1%, andsterilization is nearly 100% effective. Birth control pills arecombinations of a synthetic estrogen and a synthetic progestin(progesterone-like hormone). These two hormones act by negative feedbackto stop the release of GnRH by the hypothalamus and FSH (an estrogeneffect) and LH (a progestin effect) by the pituitary. By blocking LHrelease, the progestin prevents ovulation. As a backup measure, theestrogen inhibits FSH secretion so no follicles develop. Chemicalcontraception has been the center of much debate, particularly becauseof the long-term side effects of the estrogens. No solid evidence existsfor cancers caused by the pill, but cardiovascular problems are a majorconcern. Birth control pills have been implicated in blood clotting,atherosclerosis, and heart attacks. Smoking while using chemicalcontraception increases the risk of mortality tenfold or more. Campbell,N.; Biology, 2nd Ed., Benjamin/Cummings Publ., Redwood City, La., 1990.

Accordingly, there is a current need for inhibitors of MMPs. Suchinhibitors would be useful to treat or prevent cancer, tumor metastasis,angiogenesis in tumors, contraception, arthritis and connective tissuediseases, cardiovascular disease, inflammation or autoimmune diseases.Preferred inhibitors may exhibit selectivity for one or more specificMMPs than known competitive inhibitors. In addition, additional methodsthat prevent the release of gametes are needed. Suth methods willpreferably not include negative long-term side-effects.

SUMMARY OF THE INVENTION

The present invention provides compounds that inhibit MMPs. Accordingly,there is provided a compound of the invention which is a compound offormula (I):

wherein

A—X—M is a hydrophobic group;

D is O, S, (C₁-C₆)alkyl, a direct bond, SO₂, SO, C(═O)NR, C(═O)O,NRC(═O), or OC(═O);

E is a direct bond, (C₁-C₆)alkyl, (C₃-C₈)cycloalkyl, (C₂-C₆)alkenyl, or(C₂-C₆)alkynyl, wherein any alkyl, cycloalkyl, alkenyl, or alkynyl of Eis optionally substituted with one or more (C₁-C₆)alkyl, hydroxy,(C₁-C₆)alkoxy, cyano, nitro, halo, SR, NRR, or COOR, wherein each R isindependently H or (C₁-C₆)alkyl;

J is S or O;

G, T, and Q are each independently H, (C₁-C₆)alkyl, or cyano;

or a pharmaceutically acceptable salt thereof.

The present invention also provides a pharmaceutical composition thatcomprises a compound of formula (I) and a pharmaceutically acceptablecarrier.

The present invention also provides a radiolabeled compound comprising acompound of formula (I) and a radionuclide.

The present invention also provides a pharmaceutical composition thatcomprises a radiolabeled compound of formula (J) and a pharmaceuticallyacceptable carrier.

The present invention also provides a therapeutic method for preventingor treating a pathological condition or symptom in a mammal, such as ahuman, wherein the activity of an MMP is implicated and inhibition ofits action is desired, comprising administering to a mammal in need ofsuch therapy, an effective amount of a compound of formula (I), or apharmaceutically acceptable salt thereof.

The present invention also provides a method for treating or preventingcancer, angiogenesis, arthritis, connective tissue disease,cardiovascular disease, inflammation or autoimmune disease in a mammalinflicted with or at risk thereof comprising administering to the mammalin need of such treatment or prevention an effective amount of acompound of formula (I).

The present invention also provides a method for treating or preventingcancer in a mammal inflicted with or at risk thereof comprisingadministering to the mammal in need of such therapy an effective amountof a compound of formula (I), or a pharmaceutically acceptable saltthereof in conjunction with a chemotherapeutic agent, or apharmaceutically acceptable salt thereof.

The present invention also provides a method for inhibiting a matrixmetalloproteinase comprising a zinc atom, the method comprisingcontacting the matrix metalloproteinase with a compound with a groupthat can be activated for nucleophilic substitution by the zinc atom andcan form a covalent bond with a nucleophile of the matrixmetalloproteinase.

The present invention also provides a method for inhibiting a gelatinasecomprising a zinc atom, the method comprising contacting the gelatinasewith a compound with a group that can be activated for nucleophilicsubstitution by the zinc atom and can form a covalent bond with anucleophilic site of the gelatinase.

The present invention also provides a method for imaging a tumor in amammal inflicted with a tumor comprising administering to the mammal aneffective amount of a radiolabeled compound of formula (I), or apharmaceutically acceptable salt thereof, and detecting the presence ofthe radiolabeled compound.

The present invention also provides a method to image MMP activity in atumor and/or a vasculature comprising contacting the organism (e.g., invivo) with an effective amount of a compound the present invention,wherein the compound of formula (I) comprises a radionuclide; or apharmaceutically acceptable salt thereof.

The present invention also provides a method for imaging MMP activity ina tumor in a mammal inflicted with a tumor comprising administering tothe mammal in need of such imaging an effective amount of a compound thepresent invention, wherein the compound of formula (I) comprises aradionuclide; or a pharmaceutically acceptable salt thereof.

The present invention also provides a method for preventing ovulation ina mammal (e.g., human) at risk thereof comprising administering to themammal an effective amount of a compound of formula (I).

The present invention also provides a method for preventing theimplantation of a fertilized egg into the uterus of a mammal (e.g.,human) in need thereof comprising administering to the mammal aneffective amount of a compound of formula (I).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates a mechanism-based inhibition of an MMP by a compoundof the present invention.

FIG. 2 illustrates a synthesis of compounds of the present invention.

FIG. 3 illustrates a mechanism-based inhibition of an MMP by a compoundof the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The following definitions are used, unless otherwise described: halo isfluoro, chloro, bromo, or iodo. Alkyl, alkoxy, alkenyl, alkynyl, etc.denote both straight and branched groups; but reference to an individualgroup such as “propyl” embraces only the straight chain variant, abranched chain isomer such as “isopropyl” being specifically referredto. Bicyclic aryl denotes an ortho-fused bicyclic carbocyclicsubstituent having about nine to ten ring atoms in which at least onering is aromatic. Monocyclic heteroaryl encompasses a substituentattached via a ring carbon of a monocyclic aromatic ring containing fiveor six ring atoms consisting of carbon and one to four heteroatoms eachselected from the group consisting of non-peroxide oxygen, sulfur, andN(X) wherein X is absent or is H, O, (C₁-C₄)alkyl, phenyl or benzyl.Bicyclic heteroaryl encompasses a substituent of an ortho-fused bicyclicheterocycle of about eight to ten ring atoms derived therefrom,particularly a benzyl-derivative or one derived by fusing a propylene,trimethylene, or tetramethylene divalent substituent thereto. Bicyclicalkyl encompasses a substituent of an ortho-fused bicyclic alkyl ofabout eight to ten ring atoms containing five or six ring atomsconsisting of carbon and one to four ring atoms consisting ofheteroatoms selected from the group consisting of non-peroxide oxygen,sulfur, and N(X) wherein X is absent or is H, O (C₁-C₄)alkyl, phenyl orbenzyl.

It will be appreciated by those skilled in the art that compounds of theinvention having a chiral center may exist in and be isolated inoptically active and racemic forms. Some compounds may exhibitpolymorphism. It is to be understood that the present inventionencompasses any racemic, optically-active, polymorphic, orstereoisomeric form, or mixtures thereof, of a compound of theinvention, which possess the useful properties described herein, itbeing well known in the art how to prepare optically active forms (forexample, by resolution of the racemic form by recrystallizationtechniques, by synthesis from optically-active starting materials, bychiral synthesis, or by chromatographic separation using a chiralstationary phase) and how to determine MMP inhibition activity using thestandard tests described hereinbelow, or using other similar tests whichare well known in the art.

As used herein, “ovulation” is the release of an ovum from the ovarianfollicle. Stedman's Medical Dictionary, 25th Ed., Illustrated, Williams& Wilkins, Baltimore, 1990, p.1116.

As used herein, “ovum” is the female sex (reproductive) cell. Whenfertlized by a spermatozoon, an ovum is capable of developing into a newindividual of the same species. Stedman's Medical Dictionary, 25th Ed.,Illustrated, Williams & Wilkins, Baltimore, 1990, p.1116.

As used herein, “fertiliziation” is the process beginning withpenetration of the secondary oocyte by the spermatozoon and completed byinfusion of the male and female pronuclei. Stedman's Medical Dictionary,25th Ed., Illustrated, Williams & Wilkins, Baltimore, 1990, p.573.

As used herein, a “uterus” is the womb, metra, or the hollow muscularorgan in which the impregnated ovum is developed into the child.Stedman's Medical Dictionary, 25th Ed., Illustrated, Williams & Wilkins,Baltimore, 1990, pp.1677-1678.

Specific and preferred values listed below for substituents (i.e.,groups) and ranges are for illustration only; they do not exclude otherdefined values or other values within defined ranges for thesubstituents.

Specifically, (C₁-C₆)alkyl can be methyl, ethyl, propyl, isopropyl,butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl; (C₁-C₆)alkoxycan be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy,sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy; (C₂-C₆)alkenyl can bevinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,1,-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, 4-hexenyl, or 5-hexenyl; (C₂-C₆)alkynyl can be ethynyl,1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl,2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl,4-hexynyl, or 5-hexynyl; (C₁-C₆)alkanoyl can be acetyl, propanoyl orbutanoyl; (C₂-C₆)alkanoyloxy can be acetoxy, propanoyloxy, butanoyloxy,isobutanoyloxy, pentanoyloxy, or hexanoyloxy; (C₃-C₈)cycloalkyl can becyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, orcyclooctyl; aryl can be phenyl, indenyl, 5,6,7,8-tetrahydronaphthyl, ornaphthyl and heteroaryl can be furyl, imidazolyl, tetrazolyl, pyridyl,(or its N-oxide), thienyl, pyrimidinyl (or its N-oxide), indolyl, orquinolyl (or its N-oxide); bicyclic aryl can be indenyl or naphthyl;monocyclic heteroaryl can be furyl, imidazolyl, triazolyl, triazinyl,oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl, pyrrolyl,pyrazinyl, tetrazolyl, pyridyl (or its N-oxide), thienyl, or pyrimidinyl(or its N-oxide), bicyclic heteroaryl can be quinolyl (or its N-oxide);and bicyclic alkyl can be decahydroquinoline or decahydronaphthalene(cis and trans).

As used herein, an “amino acid” is a natural amino acid residue (e.g.Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Hyl, Hyp, Ile, Leu, Lys,Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D or L form, as well asunnatural amino acid (e.g. phosphoserine; phosphothreonine;phosphotyrosine; hydroxyproline; gamma-carboxyglutamate; hippuric acid;octahydroindole-2-carboxylic acid; statine;1,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid; penicillamine;ornithine; citruline; α-methyl-alanine; para-benzoylphenylalanine;phenylglycine; propargylglycine; sarcosine; and tert-butylglycine)residue having one or more open valences. The term also comprisesnatural and unnatural amino acids bearing amino protecting groups (e.g.acetyl, acyl, trifluoroacetyl, or benzyloxycarbonyl), as well as naturaland unnatural amino acids protected at carboxy with protecting groups(e.g. as a (C₁-C₆)alkyl, phenyl or benzyl ester or amide). Othersuitable amino and carboxy protecting groups are known to those skilledin the art (See for example, T. W. Greene, Protecting Groups In OrganicSynthesis; Wiley: New York, 1981; D. Voet, Biochemistry, Wiley: NewYork, 1990; L. Stryer, Biochemistry, (3rd Ed.), W.H. Freeman and Co.:New York, 1975; J. March, Advanced Organic Chemistry Reactions,Mechanisms and Structure, (2nd Ed.), McGraw Hill: New York, 1977; F.Carey and R. Sundberg, Advanced Organic Chemistry, Part B: Reactions andSynthesis, (2nd Ed.), Plenum: New York, 1977; and references citedtherein). According to the invention, the amino or carboxy protectinggroup can also comprise a radionuclide (e.g., Fluorine-18, Iodine-123,or Iodine-124).

As used herein, an “electrophile” refers to a chemical species, ion, ora portion of a compound which, in the course of a chemical reaction,will acquire electrons, or share electrons, to form other molecules orions. Electrophiles are ordinarily thought of as cationic species(positively charged). McGraw-Hill Concise Encyclopedia of Science &Technology, McGraw-Hill, p.715, 4^(th) Edition, NY, N.Y. (1998).

As used herein, a “nucleophile” refers to a chemical species, ion, or aportion of a compound which, in the course of a chemical reaction, willlose electrons, or share electrons, to form other molecules or ions.Nucleophiles are ordinarily thought of as anionic species (negativelycharged). Typical nucleoplic species include, e.g., hydroxyl (OH), halo(F, Cl, Br, or I), cyano (CN), alkoxy (CH₃CH₂O), carboxyl (COO), andthio (S). McGraw-Hill Concise Encyclopedia of Science & Technology,McGraw-Hill, p.715, 4th Edition, NY, N.Y. (1998).

As used herein, a “peptide” is a sequence of 2 to 25 amino acids (e.g.as defined hereinabove) or peptidic residues having one or more openvalences. The sequence may be linear or cyclic. For example, a cyclicpeptide can be prepared or may result from the formation of disulfidebridges between two cysteine residues in a sequence. A peptide can belinked through the carboxy terminus, the amino terminus, or through anyother convenient point of attachment, such as, for example, through thesulfur of a cysteine. Peptide derivatives can be prepared as disclosedin U.S. Pat. Nos. 4,612,302; 4,853,371; and 4,684,620. Peptide sequencesspecifically recited herein are written with the amino terminus on theleft and the carboxy terminus on the right.

As used herein, a “hydrophobic group” or “hydrophobic moiety” refers toa group that is relatively non-polar and will have a relatively minimalaffinity for water. The nature of the hydrphobic group (i.e., A—X—M) isnot important, provided the hydrophobic group fits into the pocket andhas a favorable interaction (e.g., binding) with the enzyme. Thehydrophobic group, while being relatively hydrophobic, can include oneor more heteroatoms (e.g., S, O, or N) that can have an electrostaticcharge or can include one or more groups (e.g., esters or amides) thatcan have an electrostatic charge, provided the hydrophobic group fitsinto the pocket and has a favorable interaction with the enzyme.

Any suitable hydrophobic group can be employed as A—X—M, provided thehydrophobic group fits into the pocket and has a favorable interaction(e.g., binding) with the enzyme. For example, the hydrophobic group caninclude a straight-chained or branched hydrocarbon chain (e.g., alkyl,alkenyl, or alkynyl), an aryl group (e.g., monocyclic or bicylic), aheteroaryl group (e.g., monocyclic or bicylic), a cycloalkyl group, anamino acid, a peptide, or a combination thereof.

In one embodiment, A—X—M can be a saturated or partially unsaturatedhydrocarbon chain comprising one or more carbon atoms and optionallycomprising one or more oxy (—O—), thio (—S—), sulfinyl (—SO—), sulfonyl(S(O)₂—), or NR_(f) in the chain, wherein each Rf is independentlyhydrogen or (C₁-C₆)alkyl. The saturated or partially unsaturatedhydrocarbon chain can optionally be substituted with one or more oxo(═O), hydroxy, cyano, halo, nitro, trifluoromethyl, trifluoromethoxy,(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)alkoxy(C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, aryl, heteroaryl, (C₃-C₈)cycloalkyl(C₁-C₆)alkyl,(aryl)(C₁-C₈)alkyl, (heteroaryl)(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl oxy,(aryl)oxy, (heteroaryl)oxy, (C₃-C₈)cycloalkyl, (aryl)oxy(aryl),(heteroaryl)oxy(heteroaryl), (C₃-C₈)cycloalkyl oxy (C₁-C₆)alkyl,(aryl)oxy (C₁-C₆)alkyl, or (heteroaryl)oxy (C₁-C₆)alkyl. In addition,any aryl, (C₃-C₈)cycloalkyl, or heteroaryl can optionally be substitutedwith one or more oxo (═O), hydroxy, cyano, halo, nitro, trifluoromethyl,trifluoromethoxy, (C₁-C₆)alkyl, (C₁-C₆)alkoxy,(C₁-C₆)alkoxy(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl, aryl, heteroaryl,(C₃-C₈)cycloalkyl(C₁-C₆)alkyl, (aryl)(C₁-C₈)alkyl,(heteroaryl)(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl oxy, (aryl)oxy,(heteroaryl)oxy, (C₃-C₈)cycloalkyl, (aryl)oxy(aryl),(heteroaryl)oxy(heteroaryl), (C₃-C₈)cycloalkyl oxy (C₁-C₆)alkyl,(aryl)oxy (C₁-C₆)alkyl, or (heteroaryl)oxy (C₁-C₆)alkyl.

When A—X—M is a “partially unsaturated” group, such group may compriseone or more (e.g. 1 or 2) carbon—carbon double or triple bonds. Forexample, when A—X—M is a partially unsaturated (C₁-C₆)alkyl, it can bevinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl,1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 2,4-hexadienyl, 5-hexenyl,ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl,1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 5-hexene-1-ynyl,2-hexynyl, 3-hexynyl, 3-hexen-5-ynyl, 4-hexynyl, or 5-hexynyl.

A specific value for A—X—M is A and M are each independently phenyl ormonocyclic heteroaryl, wherein any phenyl or heteroaryl is optionallysubstituted with one or more (e.g., 1, 2, 3, or 4) hydroxy,(C₁-C₆)alkyl, (C₁-C₆)alkanoyl, (C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxy, cyano,nitro, halo, trifluoromethyl, trifluoromethoxy, SR, NRR, or COOR; and

X is O S, SO, SO₂, C(═O)NR, C(═O)O, NRC(═O), OC(═O), NR, a direct bond,or (C₁-C₆)alkyl optionally substituted with one or more hydroxy,(C₁-C₆)alkoxy, cyano, nitro, halo, SR, NRR, or COOR.

Another specific value for A—X—M is bicyclic aryl (e.g., naphthyl),bicyclic heteroaryl, or bicyclic alkyl; wherein any aryl, heteroaryl oralkyl is optionally substituted with one or more (e.g., 1, 2, 3, or 4)hydroxy, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl, (C₁-C₆)alkanoyloxy,(C₁-C₆)alkoxy, cyano, nitro, halo, trifluoromethyl, trifluoromethoxy,SR, NRR, or COOR;

wherein each R is independently H, (C₁-C₆)alkyl, phenyl, benzyl, orphenethyl.

A specific value for A is phenyl or monocyclic heteroaryl. Anotherspecific value for A is phenyl.

A specific value for M is phenyl or monocyclic heteroaryl. Anotherspecific value for M is phenyl.

A specific value for X is O, S, SO, SO₂, C(═O)NR, C(═O)O, NRC(═O),OC(═O), NR, a direct bond, or (C₁-C₆)alkyl. Another specific value for Xis O.

Another specific value for A—X—M is:

wherein

X′ is O, (C₁-C₆)alkyl (e.g., CH₂), or a direct bond;

Y′ is N or (C₁-C₆)alkyl (e.g., CH₂); and

Z′ is halo, (C₁-C₆)alkoxy (e.g., OCH₃), or hydroxy.

Another specific value for A—X—M is:

wherein

each W′ is independently N or CH; and

Z′ is halo, (C₁-C₆)alkoxy (e.g., OCH₃), or hydroxy.

Another specific value for A—X—M is:

wherein

n′ is about 1 to about 4; and

Z′ is halo, (C₁-C₆)alkoxy (e.g., OCH₃), or hydroxy.

Another specific value for A—X—M is:

wherein

R′ is O, (C₁-C₆)alkyl (e.g., CH₂), or S; and

m′ is about 2 to about 7.

Another specific value for A—X—M is:

wherein

n′ is about 1 to about 4.

Another specific value for A—X—M is:

wherein

R′ is O, CH₂, or S.

A specific value for D is SO₂.

A specific value for E is (C₁-C₆)alkyl. Another specific value for E ismethyl.

A specific value for (C₁-C₆)alkyl is methyl.

A specific value for J is S.

A specific value for G is hydrogen.

A specific value for T is hydrogen.

A specific value for Q is hydrogen.

A specific compound of the present invention is a compound of formula(I) wherein A is phenyl, M is phenyl, X is O, D is SO₂, E is methyl, Jis S, G is hydrogen, T is hydrogen, and Q is hydrogen.

FIG. 2 illustrates a synthesis for compounds 1-4. 4-phenoxythiophenol 10was prepared from the commercially available 4-phenoxyphenol 7 via the 3step procedure illustrated by Newman and Karnes. Newman M. S.; Kames H.A. J. Org. Chem., 1996, 31, 3980-3984. Subsequent alkylation of 10 withallyl bromide, 4-bromo-1-butene and 5-bromo-1-pentene respectively, ledto the sulfanyl compounds 11-13 in good yield. Although the epoxidationof 12 and 13 with mCPBA was relatively quick, taking only 2-3 days, theformation of 11 took 7 days and required a large excess of mCPBA.Finally, the conversion of the epoxides 4-6 to their correspondingthiirane derivatives 1-3, was accomplished via the treatment of eachepoxide with ammoniumthiocyanate in THF/water. Although the thiiranes 2and 3 were isolated in high yield, 93% and 85% respectively, thiirane 1could only be recovered in a very poor (i.e., 14%) yield.

Processes for preparing compounds of formula (I) or for preparingintermediates useful for preparing compounds of formula (I) are providedas further embodiments of the invention. Intermediates useful forpreparing compounds of formula (I) are also provided as furtherembodiments of the invention.

A compound of formula (I) wherein J is S can be prepared by treating acorresponding compound of formula (I) wherein J is 0 with a suitablesulfonating reagent. See, e.g., March, Advanced Organic Chemistry,Reactions, Mechanisms and Structure, 2^(nd) Ed., 1977 and Carey &Sundberg, Advanced Organic Chemistry, Part B: Reactions, 2^(nd) Ed.,1983.

A compound of formula (I) wherein J is O can be prepared by epoxidizinga corresponding compound of formula (I) wherein the ring that includes Jis an alkene. See, e.g., March, Advanced Organic Chemistry, Reactions,Mechanisms and Structure, 2^(nd) Ed., 1977 and Carey & Sundberg,Advanced Organic Chemistry, Part B: Reactions, 2^(nd) Ed., 1983.

A compound of formula (I) wherein D is SO₂ and J is O can be prepared byoxidizing a corresponding compound of formula (I) wherein D is S. See,e.g., March, Advanced Organic Chemistry, Reactions, Mechanisms andStructure, 2^(nd) Ed., 1977 and Carey & Sundberg, Advanced OrganicChemistry, Part B: Reactions, 2^(nd) Ed., 1983.

A specific group of the compounds of the present invention, that can beactivated by zinc for nucleophilic substitution and that can form acovalent bond with a nucleophile of the matrix metalloproteinase,includes a thiirane ring. Another specific group of the compounds of thepresent invention, that can be activated by zinc for nucleophilicsubstitution and that can form a covalent bond with a nucleophile of thematrix metalloproteinase, includes an oxirane ring. In addition, aspecific nucleophile of the matrix metalloproteinase which can form acovalent bond with the group of the compounds of the present invention(e.g., thiirane or oxirane) is located at the amino acid residuecorresponding to residue 404 of the matrix metalloproteinase, whereinthe numbering is based on the active site general base for gelatinase A,which is observed in other MMPs. More specifically, the nucleophile is acarboxy (i.e., COO⁻) oxygen atom located at amino acid residuecorresponding to residue 404 of the matrix metalloproteinase, whereinthe numbering is based on the active site general base for gelatinase A,which is observed in other MMPs. See, FIG. 1.

The matrix metalloproteinase can be a human matrix metalloproteinase. Inaddition, the matrix metalloproteinase can be a gelatinase, collagenase,stromelysin, membrane-type MMP, or matrilysin. Specifically, thegelatinase can be MMP-2 or MMP-9.

According to the method of the invention, the matrix metalloproteinasecan be contacted with the compound, e.g., a compound of formula (I), invitro. Alternatively, the matrix metalloproteinase can be contacted withthe compound, e.g., a compound of formula (I), in vivo.

Without being bound by any particular theory, coordination of a thiiranein a compound of formula (I) with the enzyme active-site zinc ion isbelieved to activate the thiirane for modification by a nucleophile ofthe enzyme. See, FIG. 1. A computational model based onthree-dimensional homology modeling for this enzyme with compound 1indicates that the biphenyl group would fit in the active siteanalogously to the same group in certain known reversible inhibitors ofMMP-2 and MMP-9, as analyzed by X-ray structure determination. Freskos,J. N.; Mischke B. V.; DeCrescenzo, G. A.; Heintz, R.; Getman, D. P.;Howard, S. C.; Kishore, N. N.; McDonald, J. J.; Munie, G. E.; Rangwala,S.; Swearingen, C. A.; Voliva, C.; Welsch, D. J. Bioorg. & Med. Chem.Letters, 1999, 9, 943-948. Tamura, Y.; Watanabe, F.; Nakatani, T.;Yasui, K.; Fuji, M.; Komurasaki, T.; Tsuzuki, H.; Maekawa, R.; Yoshioka,T.; Kawada, K.; Sugita, K.; Ohtani, M. J. Med. Chem. 1998, 41, 640-649.As such, the biphenyl ether moiety in compounds 1-4 is believed to fitin the P1′ subsite of gelatinases, which is a deep hydrophobic pocket.(a) Morgunova, E.; Tuuttila, A.; Bergmann, U.; Isupov, M.; Lindqvist,Y.; Schneider, G.; Tryggvason, K. Science 1999, 284, 1667-1670. (b)Massova, I.; Fridman, R.; Mobashery, S. J. Mol. Mod. 1997, 3, 17-34;Olson, M. W.; Bernardo, M. M.; Pietila, M.; Gervasi, D. C.; Toth, M.;Kotra, L. P.; Massova, I.; Mobashery, S.; Fridman, R. J. Biol. Chem.,2000, 275, 2661-2668. This binding mode brings the sulfur of thethiirane in 1 into the coordination sphere of the zinc ion. See, FIG. 1.The models also indicated that the thiirane moiety in compounds 2 and 3,with longer carbon backbones, would not be able to coordinate with thezinc ion as well as compound 1, but would fit in an extendedconfiguration in the active site.

It is believed that the high specificity of certain compounds of theinvention for a targeted enzyme arises predominantly from three factors.(i) the compounds satisfy the binding specificity requirements at theactive site. In this respect these compounds are not any different fromconventional reversible or affinity inhibitors. (ii) Furthermore, thestructural features of the inhibition should allow it to undergochemical activation by the zinc atom of the enzyme to generate anelectrophilic species within the active site. (iii) Finally, thereshould be a nucleophilic amino-acid residue in the active site, in theproper orientation, to react with the electrophilic species (e.g.,thiirane ring), resulting in irreversible enzyme inactivation.

By selecting a hydrophobic group (e.g., A—X—M) located a specificdistance from a group (e.g., D) that can bind (e.g., hydrogen bond) withone or more sites in the enzyme (e.g., amino acid residue 191 and/oramino acid residue 192, in gelatinase A), which is in turn located aspecific distance from a thiirane ring that can coordinate with theenzyme active-site zinc atom, one can prepare selective mechanism-basedinhibitors for a given MMP. See, FIG. 1.

Accordingly, preferred MMP inhibitors have a hydrophobic aryl moiety(e.g., A—X—M) that can fit in the deep hydrophobic pocket (i.e., P₁′subsite) of an MMP. In addition, preferred mechanism-based MMPinhibitors also have a thiirane ring that can coordinate with the enzymeactive-site zinc ion, and be modified by a nucleophile (e.g.,carboxylate group of amino acid residue 404 of MMP-2) in the enzymeactive site. See, FIG. 1. The preferred MMP inhibitors can optionallyinclude a second group (e.g., D) that can coordinate with one or moresites in the enzyme. Specifically, the second group can optionallyhydrogen bond to the one or two proton donors (e.g., amino acid residuecorresponding to residue 191 and/or amino acid residue corresponding toresidue 192 of MMP-2) in the enzyme active site. See, FIG. 1.

The present invention provides a method for identifying a mechanisticbased MMP inhibitor. The method includes providing a compound wherein(1) a hydrophobic moiety of the compound fits into a hydrophobic pocketof the MMP; (2) the compound has one or two groups that can hydrogenbond with one or two hydrogen donors of the MMP, wherein the hydrogendonors of the MMP are located at amino acid residue corresponding toresidue 191 and amino acid residue corresponding to residue 192 ofMMP-2; (3) the compound has an electrophilic group that can covalentlybond with a nucleophile of the MMP, wherein the nucleophile of the MMPis located at amino acid residue corresponding to residue 404 of MMP-2;and/or (4) the compound includes a group that can coordinate with thezinc ion of the MMP.

Preferred MMP inhibitors have a thiirane or oxirane such that the sulfuror oxygen atom of the thiirane or oxirane is located about 3 angstromsto about 4 angstroms from the zinc ion. The suitable MMP inhibitors canalso include a thiirane or oxirane ring located about 3 angstroms toabout 5 angstroms from the active site nucleophile. See, FIGS. 1 and 3.

Radiolabeled compounds of formula (I) are also useful as imaging agentsfor imaging cells comprising MMP's. Accordingly, the invention alsoprovides compounds of formula (I) that include one or more detectableradionuclides (e.g., one or more metallic radionuclide and/or one ormore non-metallic radionuclides). For example, a detectable radionuclidecan be incorporated into a compound by replacing an atom of the compoundof formula (I) with a radionuclide (e.g., non-metallic radionuclide).Alternatively, a radiolabeled compound of the invention can be preparedby linking a compound of formula (I) to a chelating group that includesa detectable radionuclide (e.g., metallic radionuclide). Such compoundscan be useful to image tissues with MMP activity or tumors, in vivo orin vitro.

As used herein, a “chelating group” is a group that can include adetectable radionuclide (e.g., a metallic radioisotope). Any suitablechelating group can be employed. Suitable chelating groups aredisclosed, e.g., in Poster Sessions, Proceedings of the 46th AnnualMeeting, J. Nuc.Med., p. 316, No. 1386; Scientific Papers, Proceedingsof the 46th Annual Meeting, J. Nuc.Med., p. 123, No. 499; ScientificPapers, Proceedings of the 46th Annual Meeting, J. Nuc.Med., p. 102, No.413; Scientific Papers, Proceedings of the 46th Annual Meeting, J.Nuc.Med., p. 102, No. 414; Scientific Papers, Proceedings of the 46thAnnual Meeting, J. Nuc.Med., p. 103, No. 415; Poster Sessions,Proceedings of the 46th Annual Meeting, J. Nuc.Med., p. 318, No. 1396;Poster Sessions, Proceedings of the 46th Annual Meeting, J. Nuc.Med., p.319, No. 1398; M. Moi et al., J. Amer. Chem., Soc., 49, 2639 (1989); S.V. Deshpande et al., J. Nucl. Med., 31, 473 (1990); G. Kuser et al.,Bioconj. Chem., 1, 345 (1990); C. J. Broan et al., J. C. S. Chem. Comm.,23, 1739 (1990); C. J. Anderson et al., J. Nucl. Med. 36, 850 (1995);U.S. Pat. No. 5,739,313; and U.S. Pat. No. 6,004,533. Specifically, thechelating group can be.

As used herein, a “detectable radionuclide” is any suitable radionuclide(i.e., radioisotope) useful in a diagnostic procedure in vivo or invitro. Suitable detectable radionuclides include metallic radionuclides(i.e., metallic radioisotopes) and non-metallic radionuclides (i.e.,non-metallic radioisotopes).

Suitable metallic radionuclides (i.e., metallic radioisotopes ormetallic paramagnetic ions) include Antimony-124, Antimony-125,Arsenic-74, Barium-103, Barium-140, Beryllium-7, Bismuth-206,Bismuth-207, Cadmium-109, Cadmium-115m, Calcium-45, Cerium-139,Cerium-141, Cerium-144, Cesium-137, Chromium-51, Cobalt-55, Cobalt-56,Cobalt-57, Cobalt-58, Cobalt-60, Cobalt-64, Copper-67, Erbium-169,Europium-152, Gallium-64, Gallium-68, Gadolinium-153, Gadolinium-157Gold-195, Gold-199, Hafnium-175, Hafnium-175-181, Holmium-166,Indium-110, Indium-111, Iridium-192, Iron-55, Iron-59, Krypton-85,Lead-210, Manganese-54, Mercury-197, Mercury-203, Molybdenum-99,Neodymium-147, Neptunium-237, Nickel-63, Niobium-95, Osmium-185+191,Palladium-103, Platinum-195m, Praseodymium-143, Promethium-147,Protactinium-233, Radium-226, Rhenium-186, Rhenium-188, Rubidium-86,Ruthenium-103, Ruthenium-106, Scandium-44, Scandium-46, Selenium-75,Silver-110m, Silver-111, Sodium-22, Strontium-85, Strontium-89,Strontium-90, Sulfur-35, Tantalum-182, Technetium-99m, Tellurium-125,Tellurium-132, Thallium-204, Thorium-228, Thorium-232, Thallium-170,Tin-113, Tin-114, Tin-117m, Titanium-44, Tungsten-185, Vanadium-48,Vanadium-49, Ytterbium-169, Yttrium-86, Yttrium-88, Yttrium-90,Yttrium-91, Zinc-65, and Zirconium-95.

Specifically, the chelating group can include more than one metallicradioisotope. More specifically, the detectable chelating group caninclude 2 to about 10, 2 to about 8, 2 to about 6, or 2 to about 4metallic radioisotopes.

Specifically, the non-metallic radionuclide can be a non-metallicparamagnetic atom (e.g., Fluorine-19); or a non-metallic positronemitting radionuclide (e.g., Carbon-11, Fluorine-18, Iodine-123, orBromine-76).

Specifically, the compounds of the present invention can include morethan one non-metallic radioisotope. More specifically, the compounds ofthe present invention can include 2 to about 10, 2 to about 8, 2 toabout 6, or 2 to about 4 non-metallic radioisotopes.

A compound of formula (I), or a pharmaceutically acceptable saltthereof, can be administered to a mammal (e.g., human) in conjunctionwith a chemotherapeutic agent, or a pharmaceutically acceptable saltthereof. Accordingly, a compounds of formula (I) can be administered inconjunction with a chemotherapeutic agent to treat a tumor or cancer.

As used herein, a “chemotherapeutic agent” is a compound that hasbiological activity against one or more forms of cancer and can beadministered to a patient with a compound of formula (I) without losingits anticancer activity. Suitable chemotherapeutic agents include, e.g.,antineoplasts. Representative antineoplasts include, e.g., adjuncts,androgen inhibitors, antibiotic derivatives, antiestrogens,antimetabolites, cytotoxic agents, hormones, immunomodulators, nitrogenmustard derivatives and steroids. Physicians' Desk Reference, 50thEdition, 1996.

Representative adjuncts include, e.g., levamisole, gallium nitrate,granisetron, sargramostim strontium-89 chloride, filgrastim,pilocarpine, dexrazoxane, and ondansetron. Physicians' Desk Reference,50th Edition, 1996.

Representative androgen inhibitors include, e.g., flutamide andleuprolide acetate. Physicians' Desk Reference, 50th Edition, 1996.

Representative antibiotic derivatives include, e.g., doxorubicin,bleomycin sulfate, daunorubicin, dactinomycin, and idarubicin.

Representative antiestrogens include, e.g., tamoxifen citrate andanalogs thereof. Physicians' Desk Reference, 50th Edition, 1996.Additional antiestrogens include nonsteroidal antiestrogens such astoremifene, droloxifene and roloxifene. Magarian et al., CurrentMedicinal Chemistry, 1994, Vol. 1, No. 1.

Representative antimetabolites include, e.g., fluorouracil, fludarabinephosphate, floxuridine, interferon alfa-2b recombinant, methotrexatesodium, plicamycin, mercaptopurine, and thioguanine. Physicians' DeskReference, 50th Edition, 1996.

Representative cytotoxic agents include, e.g., doxorubicin, carmustine[BCNU], lomustine [CCNU], cytarabine USP, cyclophosphamide, estramucinephosphate sodium, altretamine, hydroxyurea, ifosfamide, procarbazine,mitomycin, busulfan, cyclophosphamide, mitoxantrone, carboplati,cisplati, cisplatin, interferon alfa-2a recombinant, paclitaxel,teniposide, and streptozoci. Physicians' Desk Reference, 50th Edition,1996.

Representative hormones include, e.g., medroxyprogesterone acetate,estradiol, megestrol acetate, octreotide acetate, diethylstilbestroldiphosphate, testolactone, and goserelin acetate. Physicians' DeskReference, 50th Edition, 1996.

Representative immunodilators include, e.g., aldesleukin. Physicians'Desk Reference, 50th Edition, 1996.

Representative nitrogen mustard derivatives include, e.g., melphalan,chlorambucil, mechlorethamine, and thiotepa. Physicians' Desk Reference,50th Edition, 1996.

Representative steroids include, e.g., betamethasone sodium phosphateand betamethasone acetate. Physicians' Desk Reference, 50th Edition,1996.

Additional suitable chemotherapeutic agents include, e.g., alkylatingagents, antimitotic agents, plant alkaloids, biologicals, topoisomeraseI inhibitors, topoisomerase II inhibitors, synthetics, antiangiogenicdrugs, and antibodies. See, e.g., AntiCancer Agents by Mechanism,http://www.dtp.nci.nih.gov/docs/cancer/searches/standard_mechanism_list.html,Apr. 12, 1999; Approved Anti-Cancer Agents,http://www.ctep.info.nih.gov/handbook/HandBookText/fda_agen.htm, pages1-7, Jun. 18, 1999; MCMP 611 Chemotherapeutic Drugs to Know,http//www.vet.purdue.edu/depts/bms/courses/mcmp611/chrx/drg2no61.html,Jun. 24, 1999; Chemotherapy,http://www.vetmed.1su.edu/oncology/Chemotherapy.htm, Apr. 12, 1999; andAngiogenesis Inhibitors in Clinical Trials,http://www.cancertrials.nci.nih.gov/news/angio/table.html, pages 1-5,Apr. 19, 2000.

Representative alkylating agents include, e.g., asaley, AZQ, BCNU,busulfan, bisulphan, carboxyphthalatoplatinum, CBDCA, CCNU, CHIP,chlorambucil, chlorozotocin, cis-platinum, clomesone,cyanomorpholinodoxorubicin, cyclodisone, cyclophosphamide,dianhydrogalactitol, fluorodopan, hepsulfam, hycanthone, iphosphamide,melphalan, methyl CCNU, mitomycin C, mitozolamide, nitrogen mustard,PCNU, piperazine, piperazinedione, pipobroman, porfiromycin,spirohydantoin mustard, streptozotocin, teroxirone, tetraplatin,thiotepa, triethylenemelamine, uracil nitrogen mustard, and Yoshi-864.AntiCancer Agents by Mechanism,http://dtp.nci.nih.gov/docs/cancer/searches/standard_mechanismlist.html, Apr. 12, 1999.

Representative antimitotic agents include, e.g., allocolchicine,Halichondrin B, colchicine, colchicine derivatives, dolastatin 10,maytansine, rhizoxin, paclitaxel derivatives, paclitaxel,thiocolchicine, trityl cysteine, vinblastine sulfate, and vincristinesulfate. AntiCancer Agents by Mechanism,http://dtp.nci.nih.gov/docs/cancer/searches/standard_mechanism_list.html,Apr. 12, 1999.

Representative plant alkaloids include, e.g., actinomycin D, bleomycin,L-asparaginase, idarubicin, vinblastine sulfate, vincristine sulfate,mitramycin, mitomycin, daunorubicin, VP-16-213, VM-26, navelbine andtaxotere. Approved Anti-Cancer Agents,http://ctep.info.nih.gov/handbook/HandBookText/fda_agent.htm, Jun. 18,1999.

Representative biologicals include, e.g., alpha interferon, BCG, G-CSF,GM-CSF, and interleukin-2. Approved Anti-Cancer Agents,http://ctep.info.nih.gov/handbook/HandBookText/fda_agent.htm, Jun. 18,1999.

Representative antiangiogenic drugs include e.g., marimastat, AG3340,COL-3, neovastat, BMS-275291, TNP-470, thalidomide, squalamine,combretastatin A-4 prodrug, endostatin, SU5416, SU6668,interferon-alpha, anti-VEGF antibody, EMD121974, CAI, interleukin-12,and IM862. Angiogenesis Inhibitors in Clinical Trials,http://www.cancertrials.nci.nih.gov/news/angio/table.html, pages 1-5,Apr. 19, 2000.

Representative topoisomerase I inhibitors include, e.g., camptothecin,camptothecin derivatives, and morpholinodoxorubicin. AntiCancer Agentsby Mechanism,http://dtp.nci.nih.gov/docs/cancer/searches/standard_mechanism_list.html, Apr. 12, 1999.

Additional biologicals include drugs designed to inhibit tumorvascularization, which is also known as tumor angiogenesis. These drugscan be potent antiangiogenic agents. Additional biologicals includehumanized antibodies to growth factors, for example, to HER2, signalingmolecules and adhesion receptors. Additional biologicals also includetreatment with recombinant viruses and other means of gene therapydelivery, including for example, DNA, oligonucleotides, rybozymes, andliposomes.

Representative topoisomerase II inhibitors include, e.g., mitoxantron,amonafide, m-AMSA, anthrapyrazole derivatives, pyrazoloacridine,bisantrene HCL, daunorubicin, deoxydoxorubicin, menogaril, N, N-dibenzyldaunomycin, oxanthrazole, rubidazone, VM-26 and VP-16. AntiCancer Agentsby Mechanism,http://dtp.nci.nih.gov/docs/cancer/searches/standard_mechanism_list.html,Apr. 12, 1999.

Representative synthetics include, e.g., hydroxyurea, procarbazine,o,p′-DDD, dacarbazine, CCNU, BCNU, cis-diamminedichloroplatimun,mitoxantrone, CBDCA, levamisole, hexamethylmelamine, all-trans retinoicacid, gliadel and porfimer sodium. Approved Anti-Cancer Agents,http://ctep.info.nih. gov/handbook/HandBookText/fda_agen.htm, Jun. 18,1999.

In cases where compounds are sufficiently basic or acidic to form stablenontoxic acid or base salts, administration of the compounds as saltsmay be appropriate. Examples of pharmaceutically acceptable salts areorganic acid addition salts formed with acids which form a physiologicalacceptable anion, for example, tosylate, methanesulfonate, acetate,citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate, and α-glycerophosphate. Suitable inorganic salts mayalso be formed, including hydrochloride, sulfate, nitrate, bicarbonate,and carbonate salts.

Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (e.g., sodium, potassiumor lithium) or alkaline earth metal (e.g., calcium) salts of carboxylicacids can also be made.

The compounds of formula (I) can be formulated as pharmaceuticalcompositions and administered to a mammalian host, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, i.e., orally or parenterally, by intravenous,intramuscular, topical or subcutaneous routes.

Thus, the present compounds may be systemically administered, e.g.,orally, in combination with a pharmaceutically acceptable vehicle suchas an inert diluent or an assimilable edible carrier. They may beenclosed in hard or soft shell gelatin capsules, may be compressed intotablets, or may be incorporated directly with the food of the patient'sdiet. For oral therapeutic administration, the active compound may becombined with one or more excipients and used in the form of ingestibletablets, buccal tablets, troches, capsules, elixirs, suspensions,syrups, wafers, and the like. Such compositions and preparations shouldcontain at least 0.1% of active compound. The percentage of thecompositions and preparations may, of course, be varied and mayconveniently be between about 2 to about 60% of the weight of a givenunit dosage form. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage level will beobtained.

The tablets, troches, pills, capsules, and the like may also contain thefollowing: binders such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, fructose, lactose or aspartame or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring may be added. Whenthe unit dosage form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form. For instance, tablets, pills, or capsules may be coatedwith gelatin, wax, shellac or sugar and the like. A syrup or elixir maycontain the active compound, sucrose or fructose as a sweetening agent,methyl and propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. Of course, any material used in preparing anyunit dosage form should be pharmaceutically acceptable and substantiallynon-toxic in the amounts employed. In addition, the active compound maybe incorporated into sustained-release preparations and devices.

The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscontain a preservative to prevent the growth of microorganisms.

The pharmaceutical dosage forms suitable for injection or infusion caninclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (e.g., glycerol, propylene glycol, liquid polyethyleneglycols, and the like), vegetable oils, nontoxic glyceryl esters, andsuitable mixtures thereof. The proper fluidity can be maintained, forexample, by the formation of liposomes, by the maintenance of therequired particle size in the case of dispersions or by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars, buffers or sodium chloride. Prolongedabsorption of the injectable compositions can be brought about by theuse in the compositions of agents delaying absorption, for example,aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompound in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfilter sterilization. In the case of sterile powders for the preparationof sterile injectable solutions, the preferred methods of preparationare vacuum drying and the freeze drying techniques, which yield a powderof the active ingredient plus any additional desired ingredient presentin the previously sterile-filtered solutions.

For topical administration, the present compounds may be applied in pureform, i.e., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

Useful solid carriers include finely divided solids such as talc, clay,microcrystalline cellulose, silica, alumina and the like. Useful liquidcarriers include water, alcohols or glycols or water-alcohol/glycolblends, in which the present compounds can be dissolved or dispersed ateffective levels, optionally with the aid of non-toxic surfactants.Adjuvants such as fragrances and additional antimicrobial agents can beadded to optimize the properties for a given use. The resultant liquidcompositions can be applied from absorbent pads, used to impregnatebandages and other dressings, or sprayed onto the affected area usingpump-type or aerosol sprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

Examples of useful dermatological compositions which can be used todeliver the compounds of formula I to the skin are known to the art; forexample, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat.No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman(U.S. Pat. No. 4,820,508).

Useful dosages of the compounds of formula I can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949.

Generally, the concentration of the compound(s) of formula I in a liquidcomposition, such as a lotion, will be from about 0.1-25 wt-%,preferably from about 0.5-10 wt-%. The concentration in a semi-solid orsolid composition such as a gel or a powder will be about 0.1-5 wt-%,preferably about 0.5-2.5 wt-%.

The amount of the compound, or an active salt or derivative thereof,required for use in treatment will vary not only with the particularsalt selected but also with the route of administration, the nature ofthe condition being treated and the age and condition of the patient andwill be ultimately at the discretion of the attendant physician orclinician.

In general, however, a suitable dose will be in the range of from about0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of bodyweight per day, such as 3 to about 50 mg per kilogram body weight of therecipient per day, preferably in the range of 6 to 90 mg/kg/day, mostpreferably in the range of 15 to 60 mg/kg/day.

The compound is conveniently administered in unit dosage form; forexample, containing 5 to 1000 mg, conveniently 10 to 750 mg, mostconveniently, 50 to 500 mg of active ingredient per unit dosage form.

Ideally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.5 to about75 μM, preferably, about 1 to 50 μM, most preferably, about 2 to about30 μM. This may be achieved, for example, by the intravenous injectionof a 0.05 to 5% solution of the active ingredient, optionally in saline,or orally administered as a bolus containing about 1-100 mg of theactive ingredient. Desirable blood levels may be maintained bycontinuous infusion to provide about 0.01-5.0 mg/kg/hr or byintermittent infusions containing about 0.4-15 mg/kg of the activeingredient(s).

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four or more sub-doses per day. The sub-dose itself may befurther divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye.

The ability of a compound of the invention to act as an MMP inhibitormay be determined using pharmacological models which are well known tothe art, or using the methods described hereinbelow.

Fluorescence Enzymatic Activity Assays

The enzymatic activity of MMP-2, MMP-9, and MMP-7 was monitored with thefluorescence quenched substrate MOCAcPLGLA₂pr(Dnp)-AR-NH₂. Fluorescencewas measured with a Photon Technology International (PTI)spectrofluorometer interfaced to a Pentium computer, equipped with theRatioMaster™ and FeliX™ hardware and software, respectively. The cuvettecompartment was thermostated at 25.0° C. Substrate hydrolysis wasmonitored at emission and excitation wavelengths of 328 and 393 nm andexcitation and emission band passes of 1 and 3 nm, respectively.Fluorescence measurements were taken every 4 s. Less than 10% hydrolysisof the fluorogenic substrate was monitored, as described by Knight.Knight, C. G. Methods Enzymol. 1995, 248, 18-34. Stromelysin 1 enzymaticactivity was monitored using the synthetic fluorogenic substrateMOCAcRPKPVE-Nva-WRK(Dnp)-NH₂ (Peptides International, Louisville, Ky.)at excitation and emission wavelengths of 325 and 393 nm and excitationand emission band passes of 1 and 3 nm, respectively.

Enzymes and Protein Inhibitors.

Human pro-MMP-2, pro-MMP-9, TIMP-1 and TIMP-2 were expressed in HeLa S3cells infected with the appropriate recombinant vaccinia viruses andwere purified to homogeneity, as previously described. Fridman, R.;Fuerst, T. R.; Bird, R. E.; Hoyhtya, M.; Oelkuct, M.; Kraus, S.;Komarek, D.; Liotta, L. A.; Berman, M. L.; Stetler-Stevenson, W. G. J.Biol. Chem. 1992, 267, 15398-15405. Fridman, R.; Birs, R. E.; Hoyhtya,M.; Oelkuct, M.; Komarek, D.; Liang, C. M.; Berman, M. L.; Liotta, L.A.; Stetler-Stevenson, W. G.,; Fuerst, T. R. Biochem. J. 1993, 289,411-416. Pro-MMP-2, pro-MMP-9, TIMP-1 and TIMP-2 concentrations weredetermined using the extinction coefficients of 122,800, 114,360, 26,500and 39,600 M⁻¹cm⁻¹, respectively. To obtain active MMP-2, pro-MMP-2 (7.3μM) was incubated at 37° C. for 1 h with 1 mM p-aminophenylmercuricacetate (APMA) (dissolved in 200 mM Tris) in buffer C. The enzymesolution was dialyzed against buffer D at 4° C. to remove APMA. ActiveMMP-9 was obtained by incubating pro-MMP-9 (1 μM) with heat-activatedrecombinant human stromelysin 1 (68 nM) (MMP-3, generously provided byDr. Paul Cannon, Center for Bone and Joint Research, Palo Alto, Calif.)at 37° C., for 2.5 h in buffer C.

The resulting solution was subjected to gelatin-agarose chromatographyto remove stromelysin 1. MMP-9 was eluted with buffer D containing 10%DMSO and dialyzed against the same buffer without DMSO to remove theorganic solvent. Pro-MMP-2 and pro-MMP-9 activation reactions weremonitored using the fluorescence quenched substrateMOCAcPLGLA₂pr(Dnp)-AR-NH₂ (Peptides International, Louisville, Ky.), aswill be described below. The MMP-2 and MMP-9 concentrations weredetermined by titration with TIMP-1.

Kinetic Analyses.

Progress curves were obtained by adding enzyme (0.5-2 nM) to a mixtureof fluorogenic substrate (5-7 μM) and varying concentrations ofinhibitor in buffer R containing 5-15% DMSO (final volume 2 ml), inacrylic cuvettes with stirring and monitoring the increase influorescence with time for 15-30 minutes. The progress curves werenonlinear least squares fitted to Equation 1 (Muller-Steffner, H. M.,Malver, O., Hosie, L., Oppenheimer, N. J., and Schuber, F. J. Biol.Chem. 1992, 267, 9606-9611.):

F=v _(s) t+I(v _(o) −v _(s))(1−exp(−kt))/k+F ₀  (1)

where v_(o) represents the initial rate, v_(s), the steady state rate,k, the apparent first order rate constant characterizing the formationof the steady-state enzyme-inhibitor complex and F_(o), the initialfluorescence, using the program SCIENTIST (MicroMath ScientificSoftware, Salt Lake City, Utah). The obtained k values, v₀ and v_(s)were further analyzed according to Equations 2 and 3 for a one-stepassociation mechanism

k=k _(off) +k _(on) [I]/(1+[S]/Km)  (2)

(v_(o) −v _(s))/v_(s) =[I]/(K _(i)(1+[S]/K _(m)))  (3)

Intercept and slope values, obtained by linear regression of the kversus inhibitor concentration plot (Equation 2), yielded theassociation and dissociation rate constants k_(on) and k_(off),respectively, and the inhibition constant K_(i) (k_(off)/k_(on)).Alternatively, K_(i) was determined from the slope of the (v_(o) −v_(s))/v_(s)vs [I] plot according to Equation 3.

The dissociation rate constants were determined independently from theenzyme activity recovered after dilution of a pre-formedenzyme-inhibitor complex. To this end, typically 200 nM of enzyme wasincubated with 1 μM of inhibitor for a sufficient time to reachequilibrium (>45 min) at 25.0° C. The complex was diluted into 2 mL ofbuffer R containing fluorogenic substrate (5-7 μM final concentration)to a final enzyme concentration of 1 nM. Recovery of enzyme activity wasmonitored for ˜30 min. The fluorescence versus time trace was fitted,using the program SCIENTIST, to Equation 4

F=v _(s) t+(v _(o) −v _(s))(1−exp(−k _(off)))/k _(off) +F ₀  (4)

where v_(o) represents the initial rate (very small), v_(s), the rateobserved when the E.I complex is completely dissociated and k_(off), thefirst order rate constant when the E.I dissociation.

Analysis for linear competitive inhibition was performed in thefollowing manner. Initial rates were obtained by adding enzyme (0.5-2nM) to a mixture of fluorogenic substrate (5-7 μM) and varyingconcentrations of inhibitor in buffer R, containing 5-15% DMSO (finalvolume 1 mL) in semi-micro quartz cuvettes, and monitoring the increasein fluorescence with time for 5-10 minutes. The fluorescence versus timetraces were fitted by linear regression analysis using FeliX™. Theinitial rates were fitted to Equation 5 (Segel, I. H. in: EnzymeKinetics, Wiley Inc., New York, 1975, pp. 104.):

v/V _(max) =S/(K _(m)(1+I/K _(i))+S)  (5)

where v and V_(max) represent the initial and maximal velocities, S andI, the substrate and inhibitor concentrations, respectively, K_(m) theMichaelis-Menten constant for the substrate-enzyme reaction and K_(i)the inhibition constant, using the program SCIENTIST.

Inhibitors 1-4 all bind with the active site of the MMPs that were usedin the study, with K_(i) values of micromolar, or less, however, thebehavior of inhibitor 1 was very different. Inhibitor 1 showed a dualbehavior. It served as a mechanism-based inhibitor with a partitionratio of 79±10 (i.e. k_(cat)/k_(inact)) for MMP-2 and 416±63 for MMP-9.Furthermore, it also behaved as a slow-binding inhibitor, for which therate constants for the on-set of inhibition (k_(on)) and recovery ofactivity from inhibition (k_(off)) were evaluated (Table 1). It wouldappear that coordination of the thiirane with the zinc ion (as seen inenergy-minimized computational models; FIG. 1) would set in motion aconfornational change, which is presumed from the slow-binding kineticbehavior. The kinetic data fit the model for slow-binding inhibition.Morrison, J. F. Adv. Enzymol. 1988, 61, 201-301. Covalent modificationof the enzymes results from this conformational change. Inhibitor 1 wasincubated with MMP-2 to the point that less than 5% activity remained.This inhibitor-enzyme complex was dialyzed over three days, whichresulted in recovery of approximately 50% of the activity. Thisobservation is consistent with modification of the active site Glu-404(according to the numbering for human MMP-2), via the formation of anester bond, which is a relatively labile covalent linkage. Thetime-dependent loss of activity is not merely due to the slow-bindingbehavior. For instance, for a k_(off) of 2×10⁻³ s⁻¹ (the values are notvery different from one another in Table 1) the half time for recoveryof activity (t_(½)) is calculated at just under 6 min. The fact that 50%of activity still did not recover after dialysis over three daysstrongly argues for the covalency of enzyme modification.

Selectivity in inhibition of gelatinases by inhibitor 1 was observed.Its K_(i) values are 13.9±4 nM and 600±200 nM for MMP-2 and MMP-9,respectively. The corresponding K_(i) values are elevated to themicromolar range for the other MMPs, even for the case of MMP-3, whichdoes show the slow-binding, mechanism-based inhibition profile. Inaddition, the values for k_(on) are 611- and 78-fold larger for MMP-2and MMP-9, respectively, than that for MMP-3. Whereas the k_(off) valuesare more similar to one another, the value for MMP-2 is the smallest, sothe reversal of inhibition of this enzyme takes place more slowly.Collectively, these kinetic parameters demonstrate that inhibitor 1 canbe a potent and selective inhibitor for MMP-2, MMP-9, and especiallyMMP-2. It has been previously shown that two molecules of either TIMP-1or TIMP-2 (endogenous cellular protein inhibitors of MMPs) bind toactivated MMP-2 and MMP-9. Olson, M. W.; Gervasi, D. C.; Mobashery, S.;Fridman, R. J. Biol. Chem. 1997, 272, 29975. One binding event is highaffinity and would appear physiologically relevant, whereas the secondbinding event takes place with relatively lower affinity (micromolar).Olson, M. W.; Gervasi, D. C.; Mobashery, S.; Fridman, R. J. Biol. Chem.1997, 272, 29975. Inhibition of MMP-2 and MMP-9 by TIMPs also followsslow-binding kinetics. The kinetic parameters for these interactions atthe high affinity site are listed in Table 1. The kinetic parameters forthe slow-binding component of inhibition of MMP-2 and MMP-9 by inhibitor1 (K_(on) and K_(off)) approach closely the same parameters for those ofthe protein inhibitors. Olson, M. W.; Gervasi, D. C.; Mobashery, S.;Fridman, R. J. Biol. Chem. 1997, 272, 29975-29983.

Oxiranes 4-6 inhibit MMPs in a competitive manner with higher K_(i)values. There was no evidence of slow-binding behavior ortime-dependence of loss of activity with this inhibitor with any of theMMPs.

Small-molecule inhibitor 1 follows both slow-binding and mechanism-basedinhibition in its kinetic profile. This compound appears to behave verysimilarly to the endogenous cellular protein inhibitors for MMPs (TIMPs)in the slow-binding component of inhibition. Furthermore, the inhibitoralso exhibits a covalent mechanism-based behavior in inhibition of theseenzymes. The high discrimination in targeting that inhibitor 1 displays(both in affinities and the modes of inhibition) among the otherstructurally similar MMPs is noteworthy and could serve as a paradigm inthe design of inhibitors for other closely related enzymes in thefuture.

EXAMPLES

Experimental Procedures

¹H and ¹³C NMR spectra were recorded on either a Varian Gemini-300, aVarian Mercury-400 or a Varian Unity-500 spectrometer. Chemical shiftsare reported in ppm from tetramethylsilane on the δ scale. Infraredspectra were recorded on a Nicolet 680 DSP spectrophotometer. Massspectra were recorded on a Kratos MS 8ORFT spectrometer. Melting pointswere taken on an Electrothermal melting point apparatus and areuncorrected. Thin-layer chromatography was performed with Whatmanreagents 0.25 mm silica gel 60-F plates. All other reagents werepurchased from either Aldrich Chemical Company or Across Organics.

The following buffers were used in experiments with enzymes: Buffer C(50 mM HEPES at pH 7.5, 150 mM NaCl, 5 mM CaCl₂, 0.02% Brij-35); bufferR (50 mM HEPES at pH 7.5, 150 mM NaCl, 5 mM CaCl₂, 0.01% Brij-35, and 1%v/v Me₂SO) and buffer D (50 mM Tris at pH 7.5, 150 mM NaCl, 5 mM CaCl₂,and 0.02% Brij-35).

Example 1

(4-Phenoxyphenylsulfonyl)methyloxirane (4).

To compound 11 (598 mg, 2.5 mmol) in dichloromethane (10 mL), mCPBA(2.84 g, 10 mmol, Aldrich 57-86%), was slowly added. The mixture wasstirred at room temperature for 3 days, after which time a secondportion of mCPBA (2.84 g, 10 mmol) was added. The mixture was thenstirred for another 4 days, after which time the mixture was poured intoethyl acetate (200 mL), and washed with aqueous sodium thiosulfate (3×50mL, 10% w/v), aqueous sodium bicarbonate (3×50 ml, 5% w/v), and brine(50 ml). The organic phase was dried over magnesium sulfate and wasconcentrated to provide a yellow oil. The crude material was purified bycolumn chromatography (silica, 4:1 hexanes:ethyl acetate) to givecompound 4 as a pale yellow semi-solid (501 mg, 70%). ¹H NMR (500 MHz,CDCl₃) δ 7.90-7.86 (m, 2H), 7.46-7.40 (m, 2H), 7.26-7.22 (m, 1H),7.10-6.96 (m, 4H), 3.34-3.24 (m, 2H), 2.84-2.80 (m, 1H), 2.49-2.46 (m,1H); ¹³C NMR (125 MHz, CDCl₃) δ 163.15, 154.95, 130.76, 130.51, 125.52,120.77, 117.83, 59.89, 46.13; IR(film) 3054 (w), 2919 (w), 1576 (s),1492 (s), 1320 (s), 1245 (s), 1148 (s) cm⁻¹; m/z (EI) 290 (M⁺, 100%),233 (70), 217 (50), 185 (40); HRMS (EI) calcd. for C₁₅H₁₄O₄S 290.0613,found 290.0611.

The intermediate, compound 11, was prepared as follows:

(A.) O-4-Phenoxyphenyl-N,N-dimethylthiocarbamate (8).

To a solution of 4-phenoxyphenol (7, 8.46 g, 45 mmol) in DMF (40 mL) at10° C., sodium hydride (1.83 g, 45 mmol, 60% dispersion in mineral oil)was added in small portions. After the evolution of hydrogen ceased,N,N-dimethylthiocarbamoyl chloride (6.16 g, 50 mmol) was added in oneportion. The reaction mixture was then stirred at 70° C. for 2 hours.The mixture was cooled to room temperature, poured into water (100 mL)and extracted with chloroform (3×50 mL). The combined organic extractswere washed with aqueous potassium hydroxide (50 mL, 5% w/v), and brine(10×50 mL). The organic extract was dried over magnesium sulfate andconcentrated to obtain a yellow oil. The crude material was purified bycolumn chromatography (silica, 5:1 hexanes:ethyl acetate) to givecompound 8 as a white solid (11.16 g, 90%). m.p. 50-51° C.; ¹H NMR (300MHz, CDCl₃) δ 7.38-7.31 (m, 2H), 7.14-7.08 (m, 1H), 7.06-7.00 (m, 6H),3.46 (s, 3H), 3.34 (s, 3H); ¹³C NMR (75 MHz, CDCl₃) δ 188.17, 157.26,155.16, 149.62, 130.05, 124.11, 123.71, 119.31, 43.57, 38.96; IR (KBr)3040 (m), 2938 (s), 1587 (s), 1487 (s), 1394 (s), 1287 (s), 1190 (s)cm⁻¹; m/z (EI) 273 (M⁺, 15%), 186 (100); HRMS (EI) calcd. for C₁₅H,₅NO₂S273.0823, found 273.0824.

(B.) S-4-Phenoxyphenyl-N,N-dimethylthiocarbamate (9).

Compound 8 (3.99 g, 15 mmol) was heated under argon at 260° C. for 3.5hours. The resulting dark brown oil was purified by columnchromatography using a gradient eluent system (silica, 19:1 then 9:1then 3:1 hexanes:ethyl acetate) to obtain compound 9 as a pale yellowsolid (2.55 g, 64%). m.p. 97-99° C.; ¹H NMR (400 MHz, CDCl₃) δ 7.45-7.40(m, 2H), 7.40-7.30 (m, 2H), 7.15-7.10 (m, 1H), 7.05 (d, J=8.8 Hz, 2H)6.98 (d, J=8.8 Hz, 2H) 3.08 (bs, 3H), 3.02 (bs, 3H); ¹³C NMR (100 MHz,CDCl₃) δ 167.48, 158.87, 156.53, 137.66, 130.09, 124.14, 122.39, 119.87,118.94,37.14; IR(KBr) 3037 (w), 2925 (w), 1652 (s), 1581 (s) 1486 (s),1239 (s) cm⁻¹; m/z (EI) 273 (M⁺, 25%), 257 (5), 200 (5); HRMS (EI)calcd. for C₁₅H₁₅NO₂S 273.0823, found 273.0822.

(C.) 4-Phenoxythiophenol (10).

A mixture of compound 9 (2.55 g, 9 mmol) in methanol (20 mL), andaqueous NaOH (10 mL, 10% w/v), were refluxed for 4 hours. The solutionwas cooled to room temperature and was acidified to pH 1 with aqueousHCl (1M). Water (100 mL) was added and the mixture was extracted withchloroform (3×50 mL). The combined organic extracts were washed withbrine (50 mL), dried over magnesium sulfate and concentrated to obtain ayellow oil. The crude product was purified by column chromatography(silica, 5:1 hexanes:ethyl acetate) to give compound 10 as a pale yellowoil (1.80 g, >99%). 1H NMR (300 MHz, CDCl₃) δ 7.36-7.31 (m, 2H),7.30-7.25 (m, 2H), 7.13-7.09 (m, 1H), 7.04-6.88 (m, 4H), 3.43 (s, 1H);¹³C NMR (75 MHz, CDCl₃) δ 157.30, 156.15, 132.14, 130.00, 124.04,123.95, 119.88, 119.04; IR(film) 3038 (w), 1583 (s), 1484 (s), 1236 (s),1166 (s) cm⁻¹; m/z (EI) 202 (M⁺, 100%; HRMS (EI) calcd. for C₁₂H₁₀OS202.0452, found 202.0454.

(D.) 3-(4-Phenoxyphenylsulfanyl)-1-propene (11).

To a mixture of compound 10 (516 mg, 2.7 mmol) and potassium carbonate(534 mg, 3.9 mmol) in DMF (5 mL), allyl bromide (253 μL, 2.9 mmol) wasadded in one portion. The mixture was stirred at room temperatureovernight. The crude reaction mixture was poured into ether (200 mL),washed with saturated aqueous potassium carbonate (25 mL), and brine(6×50 mL). The organic layer was dried over magnesium sulfate andconcentrated in vacuo to give a yellow oil. The crude material waspurified by column chromatography, (silica, 98:2 hexanes:ethyl acetate)to obtain the title compound as a pale yellow oil (598 mg, 93%). ¹H NMR(300 MHz, CDCl₃) δ 7.38-7.32 (m, 4H), 7.15-7.10 (m, 1H), 7.04-7.00 (m,2H), 6.97-6.92 (m, 2H), 5.92-5.82 (m, 1H), 5.10-5.04 (m, 2H), 3.50 (d,J=7.2 Hz, 2H); ¹³C NMR (75 MHz, CDCl₃) δ 157.14, 156.73, 134.01, 133.22,130.05, 129.50, 123.75, 119.40, 119.25, 117.81, 38.84; IR(film) 3078(w), 3039 (w), 1582 (s), 1484 (s), 1240 (s), 1165 (s) cm⁻¹; m/z (EI) 242(M+, 100%), 201 ([M-allyl]⁺, 100); HRMS (EI) calcd. for C₁₅H₁₄OS242.0765, found 242.0764.

Example 2

2-(4-Phenoxyphenylsulfonyl)ethyloxirane (5).

The title compound was prepared in the same manner as described for 4,with the exception that compound 12 was used in place of compound 11,and the reaction time was 2 days. The title compound was obtained as awhite solid (78%). m.p. 75-77° C.; ¹H NMR (500 MHz, CDCl₃) δ 7.84-7.80(m, 2H), 7.44-7.38 (m, 2H), 7.24-7.20 (m, 1H), 7.09-7.04 (m, 4H),3.25-3.15 (m, 2H), 3.02-2.97 (m, 1H), 2.76 (t, J=4.3 Hz, 1H), 2.49 (dd,J=3.0 and 5.0 Hz, 1H), 2.19-2.10 (m, 1H), 1.86 (m, 1H); ³C NMR (125 MHz,CDCl₃) δ 162.93, 155.02, 130.58, 130.81, 125.47, 120.69, 117.91, 53.15,50.32, 47.29, 26.23; IR(KBr disc) 3040 (s), 1580 (s), 1490 (s), 1320(s), 1248 (s), 1148 cm⁻¹; m/z (EI) 304 (M⁺, 80%), 233 (50), 217 (100);HRMS (EI) calcd. for C₁₆H₁₆O₄S 304.0769, found 304.0768.

(A.) 4-(4-Phenoxyphenylsulfanyl)-1-butene (12).

The title compound was prepared in the same manner as described for 11,with the exception that 4-bromo-1-butene was used in place of allylbromide. Compound 12 was obtained as a colorless oil (88%). ¹H NMR (400MHz, CDCl₃) δ 7.37-7.32 (m, 4H), 7.14-7.10 (m, 1H), 7.04-7.00 (m, 2H),6.96-6.88 (m, 2H), 5.90-5.80 (m, 1H), 5.12-5.02 (m, 2H), 2.98 (m, 2H),2.41-2.34 (m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 157.18, 156.50, 136.65,132.57, 130.05, 123.72, 119.55, 119.21, 116.47, 34.65, 33.71; IR(film)3076 (w), 2923 (w), 1583 (s), 1485 (s), 1239 (s) cm⁻¹; m/z (EI) 256 (M⁺,100%), 215 ([M-allyl]⁺, 90), 202 (15); HRMS (EI) calcd. for C₁₆H₁₆OS256.0922 found 256.0922.

Example 3

3-(4-Phenoxyphenylsulfonyl)propyloxirane (6).

The title compound was prepared in the same manner as described for 4,with the exception that compound 13 was used in place of compound 11,and that the reaction time was 3 days. The title compound was obtainedas a white solid (94%). ¹H NMR (500 MHz, CDCl₃) δ7.86-7.80 (m, 2H),7.44-7.39 (m, 2H), 7.25-7.22 (m, 1H), 7.10-7.04 (m, 4H), 3.21-3.08 (m,2H), 2.90-2.86 (m, 1H), 2.74 (t, J=4.5 Hz, 1H), 2.45 (dd, J=2.5 and 4.5Hz, 1H), 1.92 (quin, J=7.0 Hz, 2H), 1.85-1.78 (m, 1H), (m, 1H); ¹³C NMR(125 MHz, CDCl₃) δ 162.84, 155.08, 130.58, 130.48, 125.43, 120.70,117.88, 56.28, 51.64, 46.86, 31.17, 20.12; IR(KBr disc) 3063 (w), 2923(w), 1582 (s), 1488 (s), 1294 (s), 1246 (s), 1142 (s) cm⁻¹; m/z (EI) 318(M⁺, 40%), 290 (20), 217 (100%); HRMS (EI) calcd. for C₁₇H₁₈O₄S318.0926, found 318.0924.

(A.) 5-(4-Phenoxyphenylsulfanyl)-1-pentene (13).

The title compound was prepared in the same manner as described for 11,with the exception that 5-bromo-1-pentene was used in place of allylbromide. The title compound was obtained as a colorless oil (65%). ¹HNMR (500 MHz, CDCl₃) δ 7.37-7.34 (m, 4H), 7.13-7.09 (m, 1H), 7.03-7.00(m, 2H), 6.96-93 (m, 2H), 5.83-5.74 (m, 1H), 5.06-4.98 (m, 2H), 2.88 (t,J=7.0 Hz, 2H), 2.22-2.16 (m, 2H), 1.73 (q, J=7.0 Hz, 2H); ¹³C NMR (125MHz, CDCl₃) δ 157.23, 156.36, 137.84, 132.30, 130.41, 130.03, 123.67,119.55, 119.16, 115.62, 34.61, 32.86, 28.61; IR(film) 3075 (w), 2929(m), 1583 (s), 1484 (s), 1236 (s) cm⁻¹; m/z (EI) 270 (M⁺, 100%), 215(70), 202 (60); HRMS (EI) calcd. for C₁₇H₁₈OS 270.1078, found 270.1076.

Example 4

(4-Phenoxyphenylsulfonyl)methylthiirane (1).

To a solution of compound 4 (710 mg, 2.5 mmol) in THF (5 mL), a solutionof ammonium thiocyanate (559 mg, 7.4 mmol) in water (3 mL) was added.The reaction was stirred at room temperature for 16 hours, after whichtime it was poured into ethyl acetate (100 mL), and then washed withwater (25 mL), followed by brine (25 mL). The organic phase was driedover magnesium sulfate and was concentrated to give a white oil. Thecrude material was purified by column chromatography (silica, 8:1hexanes:ethyl acetate) to obtain compound 1 as a white solid (102 mg,14%). m.p. 99-101° C.; ¹H NMR (500 MHz, CDCl₃) δ 7.89-7.84 (m, 2H),7.46-7.40 (m, 2H), 7.26-7.22 (m, 1H), 7.11-6.96 (m, 4H), 3.52 (dd, J=5.5and 14.5 Hz, 1H), 3.17 (dd, J=7.5 and 14.5 Hz, 1H), 3.09-3.03 (m, 1H),2.53 (dd, J=2.0 and 6.0 Hz, 1H) 2.16 (dd, J=2.0 and 5.0 Hz, 1H); ¹³C NMR(125 MHz, CDCl₃) δ 163.20, 155.02, 132.13, 130.95, 130.52, 125.52,120.69, 117.97, 62.90, 26.31, 24.47; IR(KBr disc) 3030 (w), 1583 (s),1486 (s), 1317 (s), 1246 (s), 1141 (s) cm⁻¹; m/z (EI) 306 (M⁺, 2%), 242([M-SO₂]⁺, 35); HRMS (EI) calcd. for C₁₅H₁₄O₃S₂ 306.0384, found306.0382.

Example 5

2-(4-Phenoxyphenylsulfonyl)ethylthiirane (2).

The title compound was prepared in the same manner as described for 1,with the exception that compound 5 was used in place of compound 4. Thecrude material was purified by column chromatography (silica, 2:1hexanes:ethyl acetate) to give the title compound as a white solid(93%). m.p. 99-101° C.; ¹H NMR (500 MHz, CDCl₃) 87.83 (d, J=8.0 Hz, 2H),7.42 (t, J=8.0 Hz, 2H), 7.26-7.22 (m, 1H), 7.10-7.06 (m, 4H), 3.30-3-20(m, 2H), 2.98-2.92 (m, 1H), 2.52 (dd, J=1 and 6 Hz, 1H), 2.48-2.39 (m,1H), 2.18 (dd, J=1 and 5 Hz, 1H), 1.78-1.69 (m, 1H); ¹³C NMR (125 MHz,CDCl₃) δ 162.94, 155.03, 132.50, 130.55, 130.51, 125.48, 120.71, 117.92,55.97, 33.62, 29.82, 26.05; IR(KBr disc) 3040 (w), 1583 (s), 1487 (s),1256 (s), 1142 (s) cm⁻¹; m/z (EI) 320 (M⁺, 50%), 288,(20), 234 (40), 217(60), 170 (100); HRMS (EI) calcd. for C₁₆H₁₆O₃S₂ 320.0541, found320.0540.

Example 6

3-(4-Phenoxyphenylsulfonyl)propylthiirane (3).

The title compound was prepared in the same manner as described for 1,with the exception that compound 6 was used in place of compound 4. Thecrude material was purified by column chromatography (silica, 2:1hexanes:ethyl acetate) to give the title compound as a white solid(85%). m.p. 75-76° C.; ¹H NMR (500 MHz, CDCl₃) δ 7.85-7.82 (m, 2H),7.44-7.40 (m, 2H), 7.26-7.22 (m, 1H), 7.10-7.06 (m, 4H), 3.20-3.09 (m,2H), 2.84-2.79 (m, 1H), 2.50 (dd, J=1 and 6 Hz, 1H), 2.14 (dd, J=1 and5.5 Hz, 1H), 2.12-2.06 (m, 1H), 1.97 (quin, J=8 Hz, 2H), 1.45-1.38 (m,1H); ¹³C NMR (125 MHz, CDCl₃) δ 162.85, 155.08, 132.55, 130.60, 130.49,125.43, 120.69, 117.91, 56.09, 35.13, 34.86, 25.72, 22.92; IR(KBr disc)3000 (w), 1583 (s), 1480 (s), 1254 (s), 1143 (s) cm⁻¹; m/z (EI) 334 (M⁺,30%), 301 (10), 234 (100), 217 (70), 170 (70); HRMS (EI) calcd. forC₁₇H₁₈O₃S₂ 334.0697, found 334.06.

Example 7

TABLE 1 Kinetics parameters for inhibition of MMPs by compounds of thepresent invention k_(on)(M⁻¹s⁻¹) × 10 ⁻⁴ k_(off)(s⁻¹) × 10³ K_(i)(μM)Compound 1 MMP-2 11 ± 1  1.5 ± 0.6a 0.0139 ± 0.0004 1.8 ± 0.1 MMP-9 1.4± 0.3  9 ± 1^(a) 0.6 ± 0.2 7.1 ± 0.5 MMP-3 (1.8 ± 0.4) ± 10⁻²  2.7 ±0.9^(a) 15 ± 6  5.5 ± 0.4 MMP-7 96 ± 41 Compound 2 MMP-2 4.7 ± 0.7 MMP-944 ± 5  MMP-3 NI^(b) MMP-7 NI MMP-1 NI Compound 3 MMP-2 4.3 ± 0.7 MMP-9181 ± 41  MMP-3 NI MMP-7 NI MMP-1 NI Compound 4 MMP-2 25 ± 2  MMP-9 186± 11  MMP-3 NI MMP-7 NI MMP-1 NI TIMP-1^(c) MMP-2 4.4 ± 0.1 1.3 ± 0.20.029 ± 0.005 MMP-9 5.2 ± 0.1 1.2 ± 0.2 0.024 ± 0.004 TIMP-2^(c) MMP-23.3 ± 0.1 0.8 ± 0.1 0.023 ± 0.004 MMP-9 2.2 ± 0.1 1.3 ± 0.2 0.058 ±0.007 Compound 5 MMP-2 5.1 ± 0.5 MMP-9 102 ± 2  MMP-3 NI^(a) MMP-7 NIMMP-1 NI Compound 6 MMP-2 10.7 ± 0.6 MMP-9 75 ± 6 MMP-3 NI^(b) MMP-7 NIMMP-1 NI ^(a)Determined by two different methods, hence the set of twonumbers. ^(b)NI, for “not inhibiting”, even at high concentrations of130-330 μM. ^(c)Kinetic parameters for the high-affinity site for TIMPswere reported earlier in Olson, M. W.; Gervasi, D. C. ; Mobashery, S.;Fridman, R., J. Biol. Chem., 1997, 272, 29975-29983.

All publications, patents, and patent documents are incorporated byreference herein, as though individually incorporated by reference. Theinvention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention. In addition, somereferences were obtained on the world wide web (www). These referencesare also incorporated by reference herein, as though individuallyincorporated by reference.

What is claimed is:
 1. A compound of formula (I):

wherein A—X—M is a hydrophobic group; D is S, SO₂, or SO, E is a directbond, (C₁-C₆)alkyl, (C₃-C₈)cycloalkyl, (C₂-C₆)alkenyl, or(C₂-C₆)alkynyl, wherein any alkyl, cycloalkyl, alkenyl, or alkynyl of Eis optionally substituted with one or more (C₁-C₆)alkyl, hydroxy,(C₁-C₆)alkoxy, cyano, nitro, halo, SR, NRR, or COOR, wherein each R isindependently H or (C₁-C₆)alkyl; J is S or O; G, T, and Q are eachindependently H, (C₁-C₆)alkyl, or cyano; or a pharmaceuticallyacceptable salt thereof.
 2. The compound of claim 1 wherein A—X—M is asaturated or partially unsaturated hydrocarbon chain comprising one ormore carbon atoms and optionally comprising one or more oxy (—O—), thio(—S—), sulfinyl (—SO—), sulfonyl (S(O)₂—), or NR_(f) in the chain,wherein each R_(f) is independently hydrogen or (C₁-C₆)alkyl; whereinthe saturated or partially unsaturated hydrocarbon chain is optionallysubstituted with one or more oxo (═O), hydroxy, cyano, halo, nitro,trifluoromethyl, trifluoromethoxy, (C₁-C₆)alkyl, (C₁-C₆)alkoxy,(C₁-C₆)alkoxy(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl, aryl, heteroaryl,(C₃-C₈)cycloalkyl(C₁-C₆)alkyl, (aryl)(C₁-C₈)alkyl(heteroaryl)(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl oxy, (aryl)oxy,(heteroaryl)oxy, (C₃-C₈)cycloalkyl, (aryl)oxy(aryl),(heteroaryl)oxy(heteroaryl), (C₃-C₈)cycloalkyl oxy (C₁-C₆)alkyl,(aryl)oxy (C₁-C₆)alkyl, or (heteroaryl)oxy (C₁-C₆)alkyl; and wherein anyaryl, (C₃-C₈)cycloalkyl, or heteroaryl is optionally substituted withone or more oxo (═O), hydroxy, cyano, halo, nitro, trifluoromethyl,trifluoromethoxy, (C₁-C₆)alkyl, (C₁-C₆)alkoxy,(C₁-C₆)alkoxy(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl, aryl, heteroaryl,(C₃-C₈)cycloalkyl(C₁-C₆)alkyl, (aryl)(C₁-C₈)alkyl,(heteroaryl)(C₁-C₆)alkyl, (C₃-C₈)cycloalkyl oxy, (aryl)oxy,(heteroaryl)oxy, (C₃-C₈)cycloalkyl, (aryl)oxy(aryl),(heteroaryl)oxy(heteroaryl), (C₃-C₈)cycloalkyl oxy (C₁-C₆)alkyl,(aryl)oxy (C₁-C₆)alkyl, or (heteroaryl)oxy (C₁-C₆)alkyl.
 3. The compoundof claim 1 wherein A and M are each independently phenyl or monocyclicheteroaryl, wherein any phenyl or monocyclic heteroaryl is optionallysubstituted with one or more hydroxy, (C₁-C₆)alkyl, (C₁-C₆)alkanoyl,(C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxy, cyano, nitro, halo, trifluoromethyl,trifluoromethoxy, SR, NRR, or COOR; and X is O, S, SO, SO₂, C(═O)NR,C(═O)O, NRC(═O), OC(═O), NR, a direct bond, or (C₁-C₆)alkyl optionallysubstituted with one or more hydroxy, (C₁-C₆)alkoxy, cyano, nitro, halo,SR, NRR, or COOR.
 4. The compound of claim 1 wherein A—X—M is bicyclicaryl, bicyclic heteroaryl, or bicyclic alkyl; wherein any aryl,heteroaryl or alkyl is optionally substituted with one or more hydroxy,(C₁-C₆)alkyl, (C₁-C₆)alkanoyl, (C₁-C₆)alkanoyloxy, (C₁-C₆)alkoxy, cyano,nitro, halo, trifluoromethyl, trifluoromethoxy, SR, NRR, or COOR;wherein each R is independently H, (C₁-C₆)alkyl, phenyl, benzyl, orphenethyl.
 5. The compound of claim 1 wherein A—X—M is bicyclic aryl,bicyclic heteroaryl, or bicyclic alkyl.
 6. The compound of claim 1wherein A—X—M is:

wherein X′ is O, CH₂, or a direct bond; Y′ is N or CH₂; and Z′ is halo,OCH₃, or hydroxy.
 7. The compound of claim 1 wherein A—X—M is:

wherein each W′ is independently N or CH; and Z′ is halo, OCH₃, orhydroxy.
 8. The compound of claim 1 wherein A—X—M is:

wherein n′ is about 1 to about 4; and Z′ is halo, OCH₃, or hydroxy. 9.The compound of claim 1 wherein A—X—M is:

wherein R′ is O, CH₂, or S; and m′ is about 2 to about
 7. 10. Thecompound of claim 1 wherein A—X—M is:

wherein n′ is about 1 to about
 4. 11. The compound of claim 1 whereinA—X—M is:

wherein R′ is O, CH₂, or S.
 12. The compound of claim 1 wherein A—X—M isnaphthyl.
 13. The compound of claim 1 wherein A is phenyl.
 14. Thecompound of claim 1 wherein M is phenyl.
 15. The compound of claim 1wherein X is O.
 16. The compound of claim 1 wherein E is (C₁-C₆)alkyl.17. The compound of claim 1 wherein E is methyl.
 18. The compound ofclaim 1 wherein J is S.
 19. The compound of claim 1 wherein G ishydrogen.
 20. The compound of claim 1 wherein T is hydrogen.
 21. Thecompound of claim 1 wherein Q is hydrogen.
 22. The compound of claim 1wherein A is phenyl, M is phenyl, X is O, D is SO₂, E is methyl, J is S,G is hydrogen, T is hydrogen, and Q is hydrogen.
 23. A pharmaceuticalcomposition comprising a compound of claim 1; and a pharmaceuticallyacceptable carrier.
 24. A radiolabeled compound comprising a compound offormula (I) as described in claim 1, and a radionuclide.
 25. Thecompound of claim 24 wherein the radionuclide is a non-metallicradionuclide.
 26. The compound of claim 25 wherein the non-metallicradionuclide is Fluortne-19, Carbon-11, Fluorine-18, Iodine-123, orBromine-76.
 27. The compound of 24 wherein the compound comprises achelating group comprising a detectable radionuclide.
 28. The compoundof claim 27 wherein the detectable radionuclide is a metallicradionuclide.
 29. The compound of claim 28 wherein the metallicradionuclide is Antimony-124, Antimony-125, Arsenic-74, Barium-103,Barium-140, Beryllium-7, Bismuth-206, Bismuth-207, Cadmium-109,Cadmium-115m, Calcium-45, Cerium-139, Cerium-141, Cerium-144,Cesium-137, Chromium-51, Cobalt-55, Cobalt-56, Cobalt-57, Cobalt-58,Cobalt-60, Cobalt-64, Copper-67, Erbium-169, Europium-152, Gallium-64,Gallium-68, Gadolinium-153, Gadolinium-157 Gold-195, Gold-199,Hafnium-175, Hafnium-175-181, Holmium-166, Indium-110, Indium-111,Iridium-192, Iron-55, Iron-59, Krypton-85, Lead-210, Manganese-54,Mercury-197, Mercury-203, Molybdenum-99, Neodymium-147, Neptunium-237,Nickel-63, Niobium-95, Osmium-185+191, Palladium-103, Platinum-195m,Praseodymium-143, Promethium-147, Protactinium-233, Radium-226,Rhenium-186, Rhenium-188, Rubidium-86, Ruthenium-103, Ruthenium-106,Scandium-44, Scandium-46, Selenium-75, Silver-110m, Silver-111,Sodium-22, Strontium-85, Strontium-89, Strontium-90, Sulfur-35,Tantalum-182, Technetium-99m, Tellurium-125, Tellurium-132,Thallium-204, Thorium-228, Thorium-232, Thallium-170, Tin-113, Tin-114,Tin-117m, Titanium-44, Tungsten-185, Vanadium-48, Vanadium-49,Ytterbium-169, Yttrium-86, Yttrium-88, Yttrium-90, Yttrium-91, Zinc-65,or Zirconium-95.
 30. A pharmaceutical composition comprising a compoundof claim 24, and a pharmaceutically acceptable carrier.